JPS62501769A - Antitumor composition containing a reactive composition of cytotoxic aldehyde and benicillamine and method of use thereof - Google Patents

Abstract

A composition for therapy of tumors having high tyrosinase activity contains the reaction product of penicillamine with a cytotoxic aldehyde, or pharmaceutically acceptable salts or esters thereof, and a pharmaceutically acceptable vehicle. The preferred compounds are 5,5-dimethyl-2-phenyl-4-thiazolidine carboxylic acid or 5,5-dimethyl-2-(2-hydroxyphenyl)-4-thiazolidine carboxylic acid, or pharmaceutically acceptable salts or esters thereof. Treatment preferably comprises an intra-arterial drip. Diagnosis of tumors suitable for treatment by means of these compositions may be accomplished by administering a radioactive copper salt and scanning for accumulations of radioactivity. This will indicate sites of high tyrosinase activity.

Description

[Detailed description of the invention] Antitumor composition containing a reaction product of a cytotoxic aldehyde and penicillamine The present invention provides improved methods and compositions for cancer treatment, and improved methods and compositions for treating cancer. and, more particularly, such compositions for tumor treatment having substantial copper concentrations. The present invention relates to methods and compositions.

Background of the invention Benzaldehyde has been shown to have antitumor activity. Kochi, M, (Ko Chi, M. et al. “Anti-tumor activity of benzaldehyde (Anti-TtlMORA)” cancer tree T, Lev.

([:ancer Treat, Rep,), 84(1) 21-3.1 , 1980.

Benzaldehyde interacts very strongly and quickly with cysteine, making it It is thought to have antitumor activity. Rapidly growing cells are normal cells. Theoretically, benzaldehyde requires more cysteine than cysteine. This treatment rapidly kills tumor cells without causing significant harm to normal cells. It can be said that it inhibits the proliferation of In addition, leukotriene-D4 has been shown to increase cell growth. It is a conjugate of arachidonic acid and cysteine, which is necessary for reproduction. . It takes cysteine from the body, which inhibits the growth of new cells. It is believed that

Other aldehydes such as salicylaldehyde, formaldehyde, and glutaraldehyde and others also have severe cytotoxicity.

Penicillamine (dimethylcysteine) is known to be therapeutically useful. There is. One property of penicillamine is that it forms a complex with copper and removes it from the body. It is important to have the ability to Therefore, penicillamine is caused by excess copper. For the treatment of diseases such as Wilson's Disease, Generally used.

It is known that certain tumors exhibit high tyrosinase activity. Tyrosinase is a metal-containing enzyme (metalloenzyme). and requires copper. Tyrosinase is used to remove copper. can be more inhibited.

Summary of the invention The purpose of the present invention is to improve the anti-tumor effect of benzaldehyde. The aim is to encourage

Another object of the invention is to provide a method suitable for anti-tumortherapy. The purpose is to alleviate the harsh effects of benzaldehyde during use.

Still another object of the present invention is to use benzaldehyde in the body to treat tumors that require treatment. The purpose is to concentrate on the body part.

A further object of the present invention is to combine benzaldehyde with penicillamine to provide the advantages of both. The purpose is to take advantage of the Ru.

Yet another object of the present invention is to provide a method for diagnosing and treating tumors with the compounds of the present invention. It is about providing.

Another object of the invention is to treat melanoma and breast cancer. To provide a method for early diagnosis of certain types of tumors including carcinoma It's there.

Yet another object of the invention is to combine salicylaldehyde with penicillamine to The object of the present invention is to provide improved anti-tumor agents for the treatment of various types of tumors.

Yet another object of the present invention is to treat cytotoxic aldehydes (cytotoxic aldehydes). ehydes) with penicillamine to treat tumors with high tyrosinase activity. The objective is to obtain antitumor agents for treatment.

These and other objects of the present invention are directed to the treatment of cytotoxic aldehydes and penicillamine. This is achieved by reacting.

For example, when benzaldehyde and penicillamine are reacted, 5,5-dimethyl -2-phenyl-4-thiazolidinecarboxylic acid is formed according to the following reaction: The pharmaceutical composition contains 5,5-dimethyl-2-phenyl-4-thiazolidi as an active ingredient. Formed on the basis of copper carboxylic acids and used for cancer treatment, especially tumors with substantial copper concentrations. It can be administered to mammals for the treatment of ulcers.

5. Remarkable therapeutic properties of 5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid The product of this reaction between benzaldehyde and penicillamine is a compound of both reactants. Possesses antitumor properties, and synergistically improves the drug efficacy of both, and eliminates inconvenient problems. It is based on the fact that excluding

Inventor's U.S. Patent Application No. 320,088, Filed No. 1981, November 1981, IO However, penicillamine alone has no resistance due to its ability to chelate with copper. Tumornases require copper for their activity. Melanoma and breast cancer Many types of tumors, such as carcinomas, have particularly high levels of It has been reported to have rosinase activity. Tyrosinase is interferon Because it has the effect of denaturing the substance, the normal production of the substance in the body is offset and its This reduces the body's ability to attack tumor cells. In addition, penicillamine It is required for the activity of tyrosinase to form chelates with copper. , essentially inactivating this tyrosinase activity; this Allows terferon to restore its action on tumor cells and also Make the administration of feron effective. On the other hand, removing tumor tyrosinase activity Without it, standard interferon cannot perform its function as expected.

Benzaldehyde reacts very strongly and quickly with cysteine. It is an effective anti-tumor compound and therefore inhibits the rapid proliferation of tumor cells. . However, benzaldehyde has a very strong effect, so Stin also produces some side effects on normal cells that contain it. Ben according to the invention The use of the reaction product of dualdehyde and penicillamine is such that the reaction product is free. Since it does not have an aldehyde group, it is not as aggressive as benzaldehyde. These drawbacks are eliminated. Also, benzaldehyde and penicillium This reaction product with penicillamine forms a chelate compound with copper. It also retains its functionality. However, there is one difference and that is the free 5.5 -dimethyl-2-phenyl-4-thiazindinecarboxylic acid and copper penicillamine The 5,5-dimethyl-2-phenyl chelate of copper is water-soluble, whereas the chelate of copper with The chelate with ru-4-thiazolidinecarboxylic acid is water-insoluble. . This difference is because the active compound precipitates at the cancer site where it chelates with copper. This is another advantage.

That is, it precipitates where it is needed. Elsewhere i.e. copper In the absence of water, it remains water-soluble and is eventually excreted from the body. It becomes roto. Tumor areas with large tyrosinase activity and therefore large amounts of copper Due to the effect of precipitation of the compound at the site, the compound directly targets tumor cells. This is the place to get it. Furthermore, the compound once undergoes a chelate reaction with copper at the cancer site. It becomes substantially more unstable and 5,5-dimethyl-2-phenyl-4-thia This results in the hydrolysis of zolidinecarboxylic acid, thereby producing benzaldehyde. liberating the hydride and then restoring its activity towards cysteine to make it most essential. ensuring its direct anti-tumor effect where it is needed. and residual penicilla Min-copper chelates become water-soluble and are removed from the water, thereby preventing cancer. Tyrosinase activity at this site is inactivated, and this inactivation is said to have an antitumor effect. Is Umono.

Another aspect of the invention is to combine benzaldehyde with other cytotoxic aldehydes or with other substances. The above-mentioned 5,5-dimethyl Can be used in the same way as thyl-2-phenyl-4-thiazolidinecarboxylic acid. This includes producing a reaction product with benicilemine that can be used.

Any aldehyde that has any toxic effect at the tumor site can be used according to the invention. can. Such aldehydes include aliphatic aldehydes such as formaldehyde. CIL%, dialdehydes such as glutaraldehyde, and phenyl groups Substituted with one or more groups such as Br%I%N02 and OH This includes aromatic aldehydes, including phenylaldehydes. These When any aldehyde is reacted with penicillamine, its aldehyde group is eliminated. The resulting compounds are effectively detoxified.

The penicillamine moiety of the reaction product reacts with copper by chelating the reaction product. , hence it has high tyrosinase activity. In other words, in the tumor site where copper is present, It will suffocate and hydrolyze, resulting in the formation of a ``magic bullet.'' It acts in the form of “c bullet Release hydride. The aldehyde itself is toxic upon contact with tumor cells; On the other hand, since benidiramine inactivates tyrosinase activity, the reaction product of the present invention It is an effective antitumor agent against tumors with substantial tyrosinase activity.

One such aldehyde is salicylaldehyde. Penicillamine as mentioned above When substituted for benzaldehyde in the reaction with 2-(2-hydroxyphenyl)-4thiazolidinecarboxylic acid is produced. child The compound also includes the 5,5-dimethyl-2-phenyl-4-thiazolidine compound described above. Similar to that of rubonic acid, it has a useful effect on tumors with high tyrosinase activity. Ru. The penicillamine portion of the molecule is used to treat cancer areas where high concentrations of copper are present. Allow to settle in minutes.

Salicylaldehyde is corrosive and is used for example against skin warts. It is easily converted to salicylic acid, which is known to be This compound is also deposited there. The compound is cytotoxic to tumor cells.

5. Described about 5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid The same dosage and usage method as for 5,5-dimethyl-2-(2-hydroxyphenyl )-4-thiazindinecarboxylic acid and penicillamine and benzaldehyde and sali Applicable to reaction products with aldehydes other than tyraldehyde.

Another aspect of the invention relates to diagnosing cancerous tumors with the compounds of the invention. . This is accomplished by administering a copper salt containing a radioactive isotope of copper.

Due to high copper concentrations at tumor sites with abnormal tyrosinase activity, radiation The copper will replace the copper already present at the site in a very short time. or by being incorporated into newly generated tyrosinase. This results in deposits at the site of the tumor. With appropriate scanning equipment, radioactive copper can be maximized. It is possible to accurately measure where the The scan shows copper at the tumor site. If it shows that the specific tumor has reached the maximum, it means that This is a clear demonstration of the scale of treatment. If there is abnormal deposit of copper in the tumor area The compound of the present invention is effective against tumors due to the presence of penicillamine, which chelates with penicillamine. It accumulates at the tumor site and exerts its effect directly at the tumor site, making it the most effective for this purpose. A suitable radioactive copper has a half-life of about 12.7 hours. It is Cu.

The radioactive substance can be administered as a water-soluble copper salt that does not exhibit any toxicity at the administered dose. blame Radioactivity scanners are now so sensitive that they can detect very small amounts of radiation. This can be done by administering radioactive copper. The preferred daily allowance of copper is 2 mg. scanning The amount of radioactive copper that needs to be administered to have the desired amount of radioactivity is 2 m It requires much less than g and can be easily achieved by standard techniques without special experiments can be measured.

This diagnostic method using radioactive copper can also be used to detect malignant tumors in the early stages of some types of tumors. Can be used to diagnose existence. Many types of tumors have very high tyrosinase activity These cancers include melanoma, breast cancer, rectal cancer, and Kin's disease, liver cancer, endometrial cancer ), laryngeal cancer, mouth Il! Includes cancer, lymphoma, tonsil cancer and neurofibrosarcoma.

Administer radioactive copper to a person predicted to have any of these 11m, and If radioactive copper is deposited in a suspected tumor, it may indicate a malignant tumor or its presence. This proves that we are positive about the existence and location of metastasis. This radioactive copper test Use may also be necessary to monitor treatment according to the invention or to continue treatment in other ways. It is effective for determining the necessity.

Detailed description of preferred embodiments Preferred compounds for use in the compositions and methods of the invention are benzaldehyde It is prepared by reacting penicillamine with penicillamine. The reaction consists of the two The reaction can be carried out in a suitable solvent for the reagents, which are preferably This is done by stirring equimolar amounts together in a solvent.

This compound and reaction is described, for example, by Nagasawa, Ei Chem, ), 18.10 47 (1981) and Vestling, M. M, ) etc., Zhu. Heterocyclic Chem, (J, Heterocy clicChem, ), 12.243 (1975). are known in the prior art. Furthermore, Besek, Jie, Jie, (Pese k J, J, 3rd class, Tetrahedron 0> (Tetrahedron), 31.907 (1975) and Tello J, Med, Chem, -C himica Therapeutica), 13149 (197B )reference. Any previous publications identifying this compound do not suggest its pharmacological properties. There is no mention of usage at all. Publications such as Terol This compound and other related compounds have been used in pharmaceutical research as radioprotectants. It has been tested for various uses. However, the publication states that 5.5-dimethyl Thyl-2-phenyl-4-thiazolidinecarboxylic acid has no use as a radioprotective agent. It states that it has no effect at all. That L.D.S. is 1200mg/g It shows something there.

A preferred method of administering the compounds of the invention is intravenously (or intraarterially) administering the alkali metal in the form of genus or ammonium salts or other water-soluble pharmaceutically acceptable salts. That's true. It can also be administered orally, intramuscularly, intravenously or rectally. Administration can also be by other suitable methods, including.

The dosage range for administration of this compound is about 25 to about 600 mg/kg 7 days. It is. If it is less than about 25 mg/kg, the effect of the compound will be minimal.

If such a minimal effect is acceptable, the use of such a small amount is It is possible. The maximum dose of 600 mg is LD. It is half of Usually, the comparison A relatively small dose is sufficient. and the preferred dose is about 33-200 mg/kg/day It is. For vascular administration, the drug is administered as a continuous infusion into the artery leading to the tumor. It is preferable to

When formulated into a pharmaceutical composition, the active ingredients include those well known in the art. It can also be mixed with pharmaceutically acceptable excipients. For example, injectable preparations If available, apply it in any of the usual sterile media that can be oil-based or water-based. It can also be prepared by forming a solution and/or suspension. Preferably, The 5,5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid is water-soluble and It is prepared in the form of an alkali metal or ammonium salt. however , the active compound as an acid or as a pharmaceutically acceptable salt or as a methyl or ethyl It can also be administered in the form of esters such as esters.

For formulation for oral administration, powders, tablets, capsules, syrups and The elixir may be prepared using any of the conventional pharmaceutical carriers well known in the art. It can also be formed using

This compound is a reaction product of penicillamine and aldehydes other than benzaldehyde. The compound according to Akira was prepared in a manner similar to that described above for benzaldehyde. It is used as a reference. Those skilled in the art will be able to ascertain this using standard laboratory techniques without undue experimentation. The exact parameters of the synthesis reaction and suitable dosages can be determined.

Example 1: Preparation of 5.5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid Construction Add 1 mole of benzaldehyde to 100 cc of 9096 ethanol with continuous stirring. dissolved in. One mole of penicillamine was dissolved in water with continued stirring. Next, the second The two solutions were mixed together with continued stirring. The solution was left at room temperature. 15 A white precipitate formed within minutes. Wash the white precipitate through a sintered glass with distilled water. It's Joshi. This washed precipitate was then dried at 100°C to obtain crystals.

Example 2: Formulation The composition for intravenous (or intra-arterial) infusion is 5,5-dimethyl in physiological saline solution at 1°C. -1 unit dosage of sodium salt of 2-phenyl-4-thiazolidinecarboxylic acid (i.e., about 33-100 mg/kg).

Example 3: Method of administration Contains radioactive copper salts in patients suspected of having tumors with substantial tyrosinase activity injection of a composition. The salt may be 64CuSO4. The dose is patient The amount should be such that it provides an amount of radioactivity that can be scanned without harm. patient However, scanning the entire body (such as a CAT scan) can detect the accumulation of radioactivity in specific areas. If the patient shows Indicates that the person is in need of treatment according to the present invention. Patient undergoes radioactive copper diagnostic test or surgically removed biopsies of the tumor for tyrosinase activity. - It is determined by other diagnostic methods such as analysis that the person is suitable for treatment according to the present invention. If found, approximately 33-100 mg/kg of 5,5-dimethyl-2-phenyl - Approximately IIl of physiological saline solution of the sodium salt of 4-thiazolidinecarboxylic acid was added to the artery. An intravenous drip is placed inside the patient, preferably through a scan of the person's entire body to determine the location of the tumor. Begin to put it into your arteries.

The administration continued daily for about a week, during which time radioactive copper was administered separately and A complete scan is performed to analyze the results of the treatment. This treatment , 37 black mice (b, 1ac k mouse) were divided into two groups. Group ■ has 18 mice. and served as a control group. Group II had 19 mice and served as the treatment group. Ta. Each mouse was inoculated subcutaneously with 100,000 mouse melanoma cells. .

After the tumor became palpable (about 1 week later), the control group IA treatment group given oral/saline 33 mg/day by oral intubation. 5.5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid of kH is given. It was. All mice in the control group died within an average of 23 days. The treatment group was 5 Death began on day 00. The last mouse in the treatment group to die was on day 700.

The treated group died at a rate of approximately 1 mouse per day from day 500 onwards. in the treatment group Tumors in mice developed more slowly than tumors in mice in the control group . Two mice in the treatment group recovered and survived. The tumor was found in the two horses that were cured. It was gone in Usu.

The same experiment was carried out using 5,5-dimethyl-2-phenyl-4- at a dose of 100 mg/kg. Repeated for 20 treatment groups of mice using thiazolidine carboxylic acid. .

Treatment began approximately - day after inoculation. The control group is the same as the control group from the previous experiment. He died at the same time. In the treatment group, no M tumor that could be detected by palpation was observed. IA Five of the mice left died between days 7 and 9. of these deaths The reason is not clear, but it is assumed that it was due to asphyxia due to intubation.

Various modifications can be made without departing from the scope of the invention, and It will be clear to those skilled in the art that the invention is not limited to what is stated in the specification. That is obvious. For example, tumors with high tyrosinase activity or Any tumor that is treated as treatable by the diagnostic test described in this can be treated with certain compounds. For specific doses, generally listed here However, it depends on the specific compound, formulation, or method of administration, and especially on experimentation. You can decide for certain patients not to Additionally, certain carriers and excipients may This invention does not constitute any limitation on the compounds of the present invention or their mode of action. It will be publicly known by the time the work becomes known to a person skilled in the art. .

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Claims (14)

[Claims] 1. Reaction products of benicillamine and cytotoxic aldehydes or pharmaceutically acceptable thereof an effective amount of the active ingredient, which is a salt or ester, and a pharmaceutically acceptable salt or ester; A composition for treating a tumor having high tyrosinase activity, comprising a carrier comprising: 2. The active ingredient is 5,5-dimethyl-2-phenyl-4-thiazolidinecarbohydrate. Claim 1, which is an acid, or a pharmaceutically acceptable salt or ester thereof. Composition according to. 3. The active ingredient is 5,5-dimethyl-2-(2-hydroxyphenyl)-4- A special thiazolidine carboxylic acid, or a pharmaceutically acceptable salt or ester thereof. A composition according to claim 2. 4. The aldehyde is formaldehyde, glutaraldehyde, or ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A composition according to claim 2, which is an aldehyde. However, the phenyl group is unsubstituted or substituted with one or more of OH, Cl, BI, I, or NO2 It is something that 5. Injectable solution or suspension in an oily pharmaceutically acceptable carrier Composition according to claim 2 in the form. 6. The 5,5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid is water-soluble. in the form of its alkali metal or ammonium salts in liquid and can be injected. A composition according to claim 2 in the form of a solution. 7.5,5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid or its administering to a patient a pharmaceutically acceptable salt or ester in an amount effective to inhibit tumor growth; A method of treating a tumor with high tyrosinase activity, the method comprising: 8. The 5,5-dimethyl-2-phenyl-4-thiazolidinecarboxylic acid is The compound is in the form of a pharmaceutically acceptable salt that renders it water-soluble and the activated A patent application in which a solution of a compound in physiological saline is continuously administered intravenously or intraarterially by infusion. The method described in Item 7 of the scope of the request. 9. Particularly, the patient is a human and the effective amount is 25-600 mg/kg/day. The method according to claim 7. 10. Non-toxic amounts of radioactive copper salts that contain detectable amounts of radioactive copper for high tyrosinase Administer to the patient an amount effective to leave the active site at the site with the amount of radioactive copper deposited. For patients with high tyrosinase activity consisting of scanning to detect A method of diagnosing the presence of a tumor. 11. Composition according to claim 2 in oral dosage form. 12. A set according to claim 2 in the form of a unit dose capsule or tablet. A product. 13. Claims wherein said carrier is selected to permit administration as an intravenous infusion. A composition according to paragraph 2. 14. Reaction of penicillamine with cytotoxic aldehydes other than benzaldehyde or a pharmaceutically acceptable salt or ester thereof. of tumors with high tyrosinase activity, and a pharmaceutically acceptable carrier. Therapeutic composition.